J Bacteriol Virol.  2019 Jun;49(2):69-80. 10.4167/jbv.2019.49.2.69.

Diverse Effects of Small Molecule Inhibitors on Actin Cytoskeleton Dynamics in HIV-1 Infection

• Shin Y
• Choi BS
• Kim KC
• Kim K
• Yoon CH

• KMID: 2452048
• DOI: http://doi.org/10.4167/jbv.2019.49.2.69

Abstract

The dynamics of the actin cytoskeleton plays a pivotal role in the process of cell division, the transportation of organelles, vesicle trafficking and cell movement. Human immunodeficiency virus type 1 (HIV-1) hijacks the actin dynamics network during the viral entry and migration of the pre-integration complex (PIC) into the nucleus. Actin dynamics linked to HIV-1 has emerged as a potent therapeutic target against HIV infection. Although some inhibitors have been intensely analyzed with regard to HIV-1 infection, their effects are sometimes disputed and the exact mechanisms for actin dynamics in HIV infection have not been well elucidated. In this study, the small molecules regulating HIV-1 infection from diverse inhibitors of the actin dynamic network were screened. Two compounds, including Chaetoglobosin A and CK-548, were observed to specifically bar the viral infection, while the cytochalasin family, 187-1, N-WASP inhibitor, Rho GTPase family inhibitors (EHop-016, CID44216842, and ML-141) and LIMK inhibitor (LIM domain kinase inhibitor) increased the viral infection without cytotoxicity within a range of ~ µM. However, previously known inhibitory compounds of HIV-1 infection, such as Latrunculin A, Jasplakinolide, Wiskostatin and Swinholide A, exhibited either an inhibitory effect on HIV-1 infection combined with severe cytotoxicity or showed no effects. Our data indicate that Chaetoglobosin A and CK-548 have considerable potential for development as new therapeutic drugs for the treatment of HIV infection. In addition, the newly identified roles of Cytochalasins and some inhibitors of Rho GTPase and LIMK may provide fundamental knowledge for understanding the complicated actin dynamic pathway when infected by HIV-1. Remarkably, the newly defined action modes of the inhibitors may be helpful in developing potent anti-HIV drugs that target the actin network, which are required for HIV infection.

Keyword

Actin dynamics network; Small molecule inhibitors; HIV infection; anti-HIV effect

MeSH Terms

Actin Cytoskeleton*
Actins*
Anti-HIV Agents
Cell Division
Cell Movement
Cytochalasins
GTP Phosphohydrolases
HIV Infections
HIV-1*
Humans
Organelles
Phosphotransferases
Transportation
Actins
Anti-HIV Agents
Cytochalasins
GTP Phosphohydrolases
Phosphotransferases

Figure

• 

  Figure 1 Dose-responsive effect on viral infectivity of compounds selected from primary screening 1 × 104 TZM-bl cells were cultured in a 100 µL medium for 24 h and then serial dilutions of the inhibitory (A) and inducible (B) compounds of HIV-1 infection were added to the cells. At 48 h after treatment, the activity of luciferases was determined as described in the Materials and Methods section.

• 

  Figure 2 Regulatory effect of compounds on viral productivity. The regulatory effect of inhibitory (A) and inducible (B) compounds of HIV-1 infectivity on viral productivity was assessed with persistently HIV-1-infected cells ACH2. 2 × 104 ACH2 cells were cultured in the presence of compounds at a final concentration of 6.25 µM with PMA (0.5 µg/mL)/ionomycin (1 µg/mL). At 48 h after treatment, the titration of HIV-1 particles in the supernatant was performed using an HIV-1 p24 ELISA kit. The data are presented as the mean ± SD (n = 3). *P < 0.05 untreated vs. compound-treated cells.

• 

  Figure 3 Inhibitory effect on Tat-mediated transcription. The transcriptional regulation of inhibitory (A) and inducible (B) compounds, as shown in Table 1, was assessed with transient transfection of Tat-expressing plasmid into the TZM-bl cells. 1 µg of Tat-expressing plasmid was introduced into the cells using Lipofectamin 2000. At 24 h after transfection, the luciferase activity was measured using a Bright Glo Luciferase Assay kit. The data were represented as relative luciferase activity compared with mock transfection. The data are presented as the mean ± SD (n = 3). * P < 0.05 untreated vs. compound-treated cells.

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